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. 2014 May 22;5(5):e1246. doi: 10.1038/cddis.2014.219

Figure 6.

Figure 6

WIF1 modulates the expression of miRNAs that have critical role in salivary gland stemness and oncogenesis. PA116 and CaExPA79 cells transfected with WIF1 for 48 h were used for total RNA isolation and subjected to real-time RT-PCR for pri-miR-144 (a), pri-let-7a (b), pri-miR-21 (c), pri-miR-200c and (d) miRNA expression analyses. The transcripts were normalized with U6 pri-miRNA. (e) PA116 and CaExPA79 cells were simultaneously transfected with a plasmid containing firefly luciferase gene with a complementary miR-200c binding site in the 3′ UTR, along with WIF1 plasmid and pRL-TK, to control for transfection efficiency. The luciferase activity was then determined. (f) CaExPA79 cells transfected with WIF1 for 48 h were used for total RNA isolation and subjected to real-time RT-PCR for mRNA expression of miR-200c downstream targets ZEB1, ZEB2, BMI1 and E-cadherin. The transcripts were normalized with β-actin. Data are the mean±S.D., n=3. *P<0.001